Cephalosporins

ABSTRACT

Novel cephalosporins of the formula ##STR1## in which R 1  is an aliphatic or cycloaliphatic hydrocarbyl radical of up to 6 carbon atoms which is optionally substituted by a carboxyl group, ##STR2## R 2  and R 3  each independently is an optionally substituted aliphatic or cycloaliphatic hydrocarbyl radical of up to 6 carbon atoms, but is not unsubstituted methyl, 
     or pharmaceutically acceptable salts thereof, are effective antibacterially and promote the growth of animals.

This is a division of application Ser. No. 730,985, filed May 6, 1985,now U.S. Pat. No. 4,686,216.

The invention relates to new cephalosporins, their use as medicaments,in particular in antibacterial therapy, and processes for theirpreparation.

Cephalosporins which contain a2-(2-aminothiazol-4-yl)-2-alkoxyiminoacetic acid radical as an acyl sidechain and a pyridiniummethyl radical in the 3-position are known fromEuropean Patent No. 64,740.

The present invention provides cephalosporins of the general formula Iand their pharmaceutically acceptable salts ##STR3## in which

R¹ represents a C₁ -C₆ -alkyl radical, which can be straight-chain,branched or cyclic and also unsaturated and can be substituted by acarboxyl group,

R² represents a C₁ -C₆ -alkyl radical, which can be straight-chain,branched or cyclic, unsaturated and optionally substituted, but whichcannot represent an unsubstituted methyl radical, and ##STR4##represents a radical of the formula ##STR5## wherein

R³ has the meaning given above for R².

Preferred compounds of the formula I are those

in which

R¹ and ##STR6## have the meaning given and

R² represents a C₁ -C₆ -alkyl radical, which can be straight-chain,branched or cyclic and unsaturated and which can be monosubstituted orpolysubstituted, preferably monosubstituted, by hydroxyl, carboxyl, C₁-C₄ -alkoxycarbonyl, C₁ -C₄ -alkylcarbonyl, carbamoyl, sulpho, cyano,nitro, amino, halogen, C₁ -C₄ -alkyl- and -dialkylamino, C₁ -C₄-alkylcarbonylamino, C₁ -C₄ -alkoxy, C₁ -C₄ -alkylthio, C₁ -C₄-alkylsulphinyl and C₁ -C₄ -alkylsulphonyl, with the proviso that R²cannot be an unsubstituted methyl radical.

Particularly preferred compounds are those

in which

R¹ and ##STR7## have the meaning given and

R² represents a C₁ -C₄ -alkyl radical, which can be straight-chain,branched or cyclic and unsaturated and which can be substituted byhydroxyl, carboxyl, C₁ -C₄ -alkoxy, halogen, nitro and amino, with theproviso that R² cannot be an unsubstituted methyl radical.

Very particularly preferred compounds of the formula I are those

in which

R¹ represents a C₁ -C₃ -alkyl radical, preferably methyl,

R² represents an optionally substituted C₁ -C₄ -alkyl radical, such as,in particular, ethyl, propyl, isopropyl, cyclopropyl, hydroxyethyl,chloroethyl, methoxymethyl, methoxyethyl, vinyl, allyl, carboxyethyl oraminoethyl, but not unsubstituted methyl, and ##STR8## has theabovementioned meaning.

The compounds of the general formula I can be obtained by a process inwhich compounds of the general formula II ##STR9## wherein

R¹ has the abovementioned meaning, in which the amine group can beprotected or unprotected, are reacted, after activation of the carboxylgroup by conversion into a mixed anhydride, for example with ethylchloroformate or methanesulphonyl chloride, after conversion into theacid halide or after conversion into an activated ester with, forexample, N-hydroxybenzotriazole and dicyclohexylcarbodiimide, withcompounds of the general formula III ##STR10## wherein

R² and ##STR11## have the abovementioned meaning, and, if appropriate,the protective groups are then split off and the desired salts or, fromsalts, the free acids are prepared.

A large number of methods known from cephalosporin or penicillinchemistry can be used for the coupling of carboxylic acids (II) toβ-lactams of the formula (III). It has proved advantageous to activatethe carboxylic acids of the general formula (II) without theamine-protective group and then to couple the activated product withβ-lactams of the formula (III), which have been dissolved as salts withan amine. Activation with sulphonic acid derivatives of the formula (V)to anhydrides of the formula (IV) is particularly advantageous:##STR12## wherein

T represents a radical R⁴ -SO₂ -O- or halogen and

R⁴ denotes an alkyl radical which has 1-10 C atoms and can optionally besubstituted by fluorine, chlorine, CN, phenyl, alkoxycarbonyl, alkoxy oralkyl, it being possible for the latter alkyl radicals to carry 1-4 Catoms, or a phenyl radical, which can optionally be substituted byfluorine, chlorine, bromine, CN, alkyl, alkoxy, alkylthio,alkoxycarbonyl--it being possible for the latter alkyl groups to carry1-4 C atoms--nitro, trifluoromethyl and phenyl.

If R⁴ is substituted, 1-3 substituents, preferably those mentioned, arepreferably present.

R⁴ very particularly preferably represents a methyl or p-tolyl radical.

The mixed anhydrides of the formula (IV) are prepared by dissolving thecarboxylic acids of the formula (II) with 1-1.4 equivalents of an aminein a solvent and reacting them with 1 to 1.2 equivalents of a sulphonicacid derivative of the formula (V).

Suitable solvents are all the solvents which are stable under thereaction conditions, such as, for example, diethyl ether,tetrahydrofuran, acetonitrile, acetone, methylene chloride, chloroformor dimethylformamide.

Suitable amines are tertiary amines, such as, for example, triethylamineor tributylamine, and also sterically hindered secondary amines, suchas, for example, diisopropylamine.

The reactions can be carried out at temperatures between -80° C. androom temperature, low temperatures avoiding isomerization of thesubstituents on the double bond. The activation is advantageouslycarried out with Cl-SO₂ CH₃ in dimethylformamide at -40° to -60° C. inthe course of 0.2 to 24 hours, preferably 0.5 to 5 hours.

The solvents mentioned for the preparation of the compounds of theformula (IV) or water can be used to dissolve the compounds of theformula (III), and the amines mentioned in the above preparation can beused as the base.

Activation of the carboxylic acids of the general formula (II) byconversion into an activated ester with, for example,N-hydroxysuccinimide and dicyclohexylcarbodiimide or1-hydroxybenzotriazole and dicyclohexylcarbodiimide is also particularlyadvantageous.

Suitable solvents are all the solvents which are also suitable for thepreparation of anhydrides of the formula (IV).

The reactions can be carried out at temperatures between -30° C. and+100° C. Advantageously, activation is carried out with1-hydroxybenzotriazole and dicyclohexylcarbodiimide in dimethylformamideat room temperature for 2 to 6 hours, the dicyclohexylurea which hasprecipitated is then filtered off with suction and the reaction with acompound of the formula (III) in the form of a solution of its aminesalt is carried out in the course of 2 to 24 hours. The solventsmentioned for the preparation of the compounds of the formula (IV) canbe used to dissolve the compounds of the formula (III), and the aminesmentioned in the above preparation can be used as the base.

The compounds of the formula (III) are obtained by splitting off theamine-protective group R⁵ from compounds of the formula (VI). ##STR13##

R⁵ here can be either a protective group which is unstable towardsacids, such as the t-butoxycarbonyl group, or, advantageously, aprotective group which can be split off enzymatically. Preferredprotective groups which can be split off enzymatically are phenacetyl or2-thienylacetyl.

The enzymatic splitting off is carried out at room temperature in wateror a mixture of water and a polar organic solvent, such as, for example,acetonitrile or tetrahydrofuran, with immobilized penicillin G acylaseat pH 7-8, preferably at pH 7.5-7.8.

During the enzymatic splitting, the pH value is kept constant byaddition of a base, such as lithium hydroxide, sodium hydroxide,potassium hydroxide or a tertiary amine, such as, for example,triethylamine, tripropylamine, tributylamine or pyridine.

The compounds of the formula (VI) can be prepared from esters of theformula (VII) via intermediate compounds of the formula (VIII).##STR14##

In the esters of the formula (VII), X represents a leaving group, suchas mesylate, tosylate, brosylate, triflate, nonaflate, iodide, bromideor chloride, and R⁶ represents an acid-protective group which iscustomary in cephalosporin chemistry, preferably a protective groupwhich can be split off under acid conditions, such as, for example,benzhydryl, 4-methoxydiphenylmethyl or t-butyl.

The compounds of the formula (VII) are converted into the reactive freeacids of the formula (VIII) by splitting off the acid-protective groupR⁶. In the case of the preferred protective groups R⁶ which are unstabletowards acids, the protective group is split off in an organic solvent.Preferably, the benzhydryl protective group is split off in methylenechloride with trifluoroacetic acid, possibly with the addition of analkoxybenzene, preferably methoxybenzene. The elimination reaction iscarried out at -20° C. to +30° C., preferably 0° C., in the course of 5minutes to one hour, preferably in the course of 20 minutes.

The acid of the formula (VIII) can be isolated after the protectivegroup has been split off. Advantageously, however, the product is notisolated but is converted directly, without purification, into compoundsof the formula (VI). For this, the solution of (VIII) formed in thereaction (VII)→(VIII) is concentrated under mild conditions in vacuo.The crude acid which remains is taken up in an organic solvent,preferably in tetrahydrofuran, and reacted with 2-50 equivalents,preferably with 5-20 equivalents, of a tertiary amine of the formula##STR15## wherein

R² and ##STR16## have the abovementioned meaning, to give compounds ofthe formula (VI).

The reaction is carried out at temperatures between -20° C. and 40° C.,preferably at 25° C., in the course of 10 minutes to two hours,preferably in the course of 30 minutes. When the reaction has ended, theproduct can be precipitated by addition of diethyl ether. The crudeproduct thus obtained can be purified on a resin, such as Diaion HP 20or XAD 7. It is also advantageously possible to further convert thecrude product directly into compounds of the formula (III).

Alternatively, the compounds of the formula (VI) can be prepared fromacids of the formula (IX) in which R⁵ has the abovementioned meaning andR⁷ represents ##STR17## an optionally substituted alkyl or aryl radical,such as methyl, ethyl, propyl, chloromethyl, dichloromethyl,trichloromethyl, trifluoromethyl or phenyl. R⁷ very particularlypreferably represents a methyl group.

The starting compounds of the formula (IX) are suspended in a suitableorganic solvent and are dissolved by silylation to the silyl ester XI.Particularly suitable organic solvents are chloroform, methylenechloride and dichlorethane. The silylation is carried out with acustomary silylating agent, such as trimethylchlorosilane (TMCS),hexamethyldisilazane (HMDS), N,O-bis-(trimethylsilyl)acetamide (BSA),N,O-bis-(trimethylsilyl)-trifluoroacetamide (BSTFA),N-methyl-N-trimethylsilylacetamide (MSA),N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA),1,3-bis-(trimethylsilyl)urea ortrimethylsilyltrifluoromethanesulphonate. A mixture of severalsilylating agents can also be employed here.

The silylation is carried out at -30° C. to +70° C., preferably at -10°C. to +10° C., in the course of 5 minutes to 30 minutes. An excess of upto ten-fold of the silylating agent, preferably a two-fold to five-foldexcess, is advantageously employed.

The solution thus obtained of the trimethylsilyl ester of the formula(X) is reacted with one to ten equivalents, preferably with three tofour equivalents, of a trialkylsilyl iodide, particularly preferablytrimethylsilyl iodide, at -40° C. to +30° C., preferably at -10° C. to+10° C., in the course of 15 minutes to 2 hours, preferably in thecourse of 30 minutes to 1 hour, to give compounds of the formula (XI).

The compounds of the formula XI are advantageously not isolated, butreacted directly, without purification, with amines ##STR18## to givethe compounds of the formula VI.

Alternatively, the compounds of the general formula I can also beprepared by reacting compounds of the formula (XII) ##STR19## in which

R¹ and R⁷ have the abovementioned meaning, directly, without isolatingthe intermediate stages, after silylation and conversion into theiodide, with amines ##STR20## to give compounds of the formula I, in amanner analogous to that described above for the conversion of compoundsof the formula X into compounds of the formula VI.

The compounds according to the invention exhibit a powerful and broadantimicrobial activity, in particular against Gram-negative andGram-positive bacteria. These properties enable them to be used aschemotherapeutic active compounds in medicine.

The compounds according to the invention are particularly effectiveagainst bacteria and bacteria-like microorganisms. They are thereforeparticularly suitable in human and veterinary medicine for theprophylaxis and chemotherapy of local and systemic infections caused bythese pathogens.

For example, local and/or systemic diseases which are caused by thefollowing pathogens or by mixtures of the following pathogens can betreated and/or prevented: Micrococcaceae, such as Staphylococci, forexample Staphylococcus aureus, Staph. epidermidis and Staph. aerogenes,and Graffkya tetragena (Staph.=Staphylococcus); Lactobacteriaceae, suchas Streptococci, for example Streptococcus pyogenes, α- andβ-haemolyzing Streptococci, non-(γ-)-haemolyzing Streptococci, Str.viridans and Str. faecalis (Enterococci) and Dipolococcus pneumoniae(Pneumococci)(Str.=Streptococcus); Enterobacteriaceae, such asEscherichiae bacteria of the Coli group: Escherichia bacteria, forexample Escherichia coli, Enterobacter bacteria, for example E.aerogenes and E. cloacae, Klebsiella bacteria, for example K.pneumoniae, Serratia, for example Serrathia marcescens (E.=Enterobacter)(K.=Klebsiella), Proteae bacteria of the Proteus group: for exampleProteus vulgaris, Pr. morganii, Pr. rettgeri and Pr. mirabilis(Pr.=Proteus); Pseudomonadaceae, such as Pseudomonas bacteria, forexample Pseudomonas aeruginosa (Ps.=Pseudomonas); and Bacteroidaceae,such as Bacteroides bacteria, for example Bacteroides fragilis(B.=Bacteroides).

The above list of pathogens is purely illustrative and is in no way tobe interpreted as restrictive.

Examples which may be mentioned of diseases which can be prevented,alleviated and/or cured by the compounds according to the invention are:diseases of the respiratory tract and pharyngeal cavity; otitis andpharyngitis; pneumonia; peritonitis and pyelonephritis; cystitis andendocarditis; systemic infections; bronchitis; arthritis; and localinfections.

The present invention includes pharmaceutical formulations which, inaddition to non-toxic, inert, pharmaceutically suitable excipients,contain one or more compounds according to the invention or whichconsist of one or more active compounds according to the invention, aswell as processes for the preparation of these formulations.

The present invention also includes pharmaceutical formulations indosage units. This means that the formulation is in the form ofindividual parts, for example tablets, dragees, capsules, pills,suppositories and ampules, the active compound contents of whichcorrespond to a fraction or a multiple of an individual dose. The dosageunits can contain, for example, 1, 2, 3 or 4 individual doses or 1/2,1/3 or 1/4 of an individual dose. An individual dose preferably containsthe amount of active compound which is given in one administration andwhich usually corresponds to a whole, a half, one third or one quarterof a daily dose.

By non-toxic, inert, pharmaceutically suitable excipients there are tobe understood solid, semi-solid or liquid diluents, fillers andformulation auxiliaries of every kind.

Tablets, dragees, capsules, pills, granules, suppositories, solutions,suspensions and emulsions, pastes, ointments, gels, creams, lotions,powders and sprays may be mentioned as preferred pharmaceuticalformulations.

Tablets, dragees, capsules, pills and granules can contain the activecompound or compounds alongside the customary excipients, such as (a)fillers and extenders, for example starches, lactose, sucrose, glucose,mannitol and silicic acid, (b) binders, for examplecarboxymethylcellulose, alginates, gelatine and polyvinylpyrrolidone,(c) humectants, for example glycerol, (d) disintegrating agents, forexample agar-agar, calcium carbonate and sodium carbonate, (e) solutionretarders, for example paraffin and (f) absorption accelerators, forexample quaternary ammonium compounds, (g) wetting agents, for examplecetyl alcohol and glycerol monostearate, (h) adsorbents, for examplekaolin and bentonite, and (i) lubricants, for example talc, calcium andmagnesium stearate and solid polyethylene glycols, or mixtures of thesubstances listed under (a) to (i).

The tablets, dragees, capsules, pills and granules can be provided withthe usual coatings and shells, optionally containing opacifying agents,and can also be of such composition that they release the activecompound or compounds only, or preferentially, in a certain part of theintestinal tract, if appropriate in a delayed manner, examples ofembedding compositions which can be used being polymeric substances andwaxes.

The active compound or compounds, optionally together with one or moreof the abovementioned excipients, can also be in microencapsulated form.

Suppositories can contain, in addition to the active compound orcompounds, the customary water-soluble or water-insoluble excipients,for example polyethylene glycols, fats, for example cacao fat, andhigher esters (for example C₁₄ -alcohol with C₁₆ -fatty acid), ormixtures of these substances.

For parenteral administration, the solutions can also be in a sterileform which is isotonic with blood.

The therapeutically active compounds should preferably be present in theabovementioned pharmaceutical formulations in a concentration of about0.1 to 99.5% by weight, preferably about 0.5 to 95% by weight of thetotal mixture.

The abovementioned pharmaceutical formulations can also contain otherpharmaceutical active compounds, in addition to the compounds accordingto the invention.

The abovementioned pharmaceutical formulations are prepared in thecustomary manner by known methods, for example by mixing the activecompound or compounds with the excipient or excipients.

The active compounds or the pharmaceutical formulations can beadministered locally, orally, parenterally, intraperitoneally and/orrectally, preferably orally or parenterally, such as intravenously orintramuscularly.

In general, it has proved advantageous both in human medicine and inveterinary medicine to administer the active compound or compoundsaccording to the invention in total amounts of about 1 to about 1,000,preferably 1 to 200 mg/kg of body weight every 24 hours, if appropriatein the form of several individual administrations, in order to achievethe desired results. An individual administration preferably containsthe active compound or compounds according to the invention in amountsof about 1 to about 250, in particular 1 to 60 mg/kg of body weight.However, it may be necessary to deviate from the dosages mentioned, andin particular to do so as a function of the species and body weight ofthe subject to be treated, the nature and severity of the diseases, thenature of the formulation and of the administration of the medicamentand the period or interval over which administration takes place. Thus,it can in some cases be sufficient to manage with less than theabovementioned amount of active compound, while in other cases theabovementioned amount of active compound must be exceeded. Theparticular optimum dosage required and the mode of administration of theactive compounds can easily be determined by anyone skilled in the arton the basis of his expert knowledge.

In order to extend the action spectrum, the active compounds accordingto the invention can be combined with another β-lactam antibiotic oralso with aminoglycoside antibiotics, such as, for example, gentamicin,sisomicin, kanamycin, amikacin or tobramycin.

The active compounds according to the invention can be used in allfields of animal breeding as agents for promoting and acceleratinggrowth and for improving the feed utilisation of healthy and sickanimals.

The activity of the active compounds here is largely independent of thespecies and sex of the animals. The active compounds have provedparticularly valuable in the rearing and keeping of young animals andfattening animals. The following stock animals and pets may be mentionedas examples of animals for which the active compounds can be used forpromoting and accelerating growth and for improving feed utilization:warm-blooded animals, such as cattle, pigs, horses, sheep, goats, cats,dogs and rabbits; fur-bearing animals, for example mink and chinchillas;poultry, for example chicken, geese, ducks, turkeys, pigeons, parrotsand canaries, and cold-blooded animals, such as fish, for example carp,and reptiles, for example snakes.

The amounts of the active compounds which are administered to theanimals to achieve the desired effect can be varied substantiallybecause of the advantageous properties of the active compounds. It ispreferably about 0.01 to 50, in particular 0.1 to 10 mg/kg of bodyweight daily. The period of administration can be from a few hours ordays up to several years. The amount of the active compound to beadministered and the corresponsing period of administration depend, inparticular, on the species, sex, state of health and nature of housingand feeding of the animals and can easily be determined by any expert.

The active compounds are administered to the animals by the customarymethods. The nature of the administration depends, in particular, on thespecies, behaviour and state of health of the animals. Thus,administration can be effected orally or parenterally once or severaltimes daily at regular or irregular intervals. For reasons ofexpediency, oral administration, in particular in the rhythm of the foodand/or drik intake of the animals, is to be preferred in most cases.Food in the context of the present invention is to be understood as bothsolid and liquid food and also drinks and water.

The active compounds can be administered as pure substances or informulated form, that is to say as a mixture with non-toxic inertcarriers of any desired type, for example with carriers and informulations such as are customary in nutritive preparations.

The active compounds, optionally in formulated form, are administered ina suitable form together with pharmaceutical active compounds, mineralsalts, trace elements, vitamins, proteins, fats, colourants and/orflavour substances.

Oral administration together with the feed and/or drinking water isrecommended, the active compound being added to all or only portions ofthe feed and/or drinking water, as required.

The active compounds are formulated by customary methods by simplemixing, as a pure mixture of substances, preferably in finely dividedform or in formulated form in a mixture with edible non-toxic carriers,if appropriate in the form of a premix or a feed concentrate, to whichthe feed and/or drinking water is added.

The feed and/or drinking water can contain, for example, the activecompounds in a weight concentration of about 0.01 to 50, in particular0.1 to 10 ppm. The optimum level of the concentration of the activecompounds in the feed and/or drinking water depends, in particular, onthe feed and/or drinking water intake of the animals and can easily bedetermined by any expert.

The nature of the feed and its composition is irrelevant here. All thecustomary or specific feed compositions which preferably contain theusual equilibrium of energy substances and builder substances necessaryfor balanced nutrition can be used. The feed can be composed, forexample, of vegetable substances, for example hay, beet, cereals andcereal by-products, animal substances, for example meat, fats and bonemeal, fish products, vitamins, for example vitamin A, D complex and Bcomplex, proteins, amino acids, for example DL-methionine, and inorganicsubstances, for example lime and sodium chloride.

Feed concentrates contain the active compounds alongside ediblesubstances, for example rye flour, maize flour, soya bean flour or lime,optionally with further nutrients and builder substances, as well asproteins, mineral salts and vitamins. They can be prepared by thecustomary methods of mixing.

In premixes and feed concentrates, preferably, the active compounds canoptionally also be protected from air, light and/or moisture by suitableagents which coat their surface, for example with non-toxic waxes orgelatine.

The following is an example of the composition of a feed for rearingchicks which contains an active compound according to the invention.

200 g of wheat, 340 g of corn, 361 g of coarse soy bean meal, 60 g ofbeef tallow, 15 g of dicalcium phosphate, 10 g of calcium carbonate, 4 gof iodinated sodium chloride, 7.5 g of a vitamin/mineral mix and 2.5 gof an active compound premix give, after thorough mixing, 1 kg of feed.

1 kg of feed mix contains: 600 I.U. of vitamin A, 100 I.U. of vitaminD₃, 10 mg of vitamin E, 1 mg of vitamin K₃, 3 mg of riboflavin, 2 mg ofpyridoxine, 20 mcg of vitamin B₁₂, 5 mg of calcium pantothenate, 30 mgof nicotinic acid, 200 mg of choline chloride, 200 mg of MnSO₄ ×H₂ O,140 mg of ZnSO₄ ×7H₂ O, 100 mg of FeSO₄ ×7H₂ O and 20 mg of CuSO₄ ×5H₂O.

The active compound premix contains the active compounds in the desiredamount, for example 10 mg, and, in addition, 1 g of DL-methionine andsoy bean flour in an amount such that 2.5 g of premix are formed.

The following is an example of the composition of a feed for rearingpigs, which contains an active compound according to the invention.

630 g of shredded cereal feed (composed of 200 g of maize, 150 g ofshredded barley, 150 g of shredded oats and 130 g of shredded wheat), 80g of fish meal, 60 g of coarse soy bean meal, 60 g of tapioca meal, 38 gof brewer's yeast, 50 g of a vitamin/mineral mix for pigs (composition,for example, as for the chick feed), 30 g of linseed cake meal, 30 g ofcorn gluten feed, 10 g of soy bean oil, 10 g of sugar cane molasses and2 g of an active compound premix (composition, for example, as for thechick feed) give, after thorough mixing, 1 kg of feed.

The feed mixtures described are preferably intended for rearing andfattening chicks and pigs, but they can also be used, in the same or asimilar composition, for rearing and fattening other animals.

EXAMPLE 1 Benzhydryl3-chloromethyl-7α-phenylacetamido-3-cephem-4-carboxylate

24 ml (0.3 mole) of pyridine, 400 ul of dimethylformamide and 21.6 ml(0.3 mole) of thionyl chloride are added to a solution of 103 g (0.2mole) of benzhydryl3-hydroxymethyl-7β-phenylacetamido-3-cephem-4-carboxylate (prepared, forexample, in accordance with Helv. Chim. Acta 57, 2044 (1974)) in 3.5 lof absolute tetrahydrofuran, while cooling with ice. After 10 minutes,the mixture is concentrated on a rotary evaporator, the residue is takenup in 2 l of ethyl acetate and the mixture is extracted by shaking twicewith sodium bicarbonate solution and once with water. The organic phaseis extracted by stirring with 50 g each of kieselguhr and activecharcoal and the extract is filtered with suction over a frit coveredwith silica gel. The filtrate is then dried over magnesium sulphate andconcentrated, the residue is taken up in 200 ml of methylene chlorideand the product is precipitated with petroleum ether.

Yield: 76 g;

¹ H-NMR (DCCl₃)δ(ppm)=7.20-7.50 (15H, m, aromatic); 6.96 (1H, s, CHO₂);6.30 (1H, d, J=9 Hz, NH); 5.86 (1H, dd, J=9 Hz, J=5 Hz, H-7); 4.95 (1H,d, J=5 Hz, H-6); 4.36 (2H, bs, DH₂ -Cl); 3.66 (1H, d, J=15 Hz, O--CH₂--); 3.58 (1H, d, J=15 Hz, O--CH₂ --); 3.56 (1H, d, J=18 Hz, H-2); and3.40 (1H, d, J=18 Hz, H-2).

EXAMPLE 27-Amino-3-(1-ethyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate

10 g (18.8 mmol) of benzhydryl3-chloromethyl-7β-phenylacetamido-3-cephem-4-carboxylate are dissolvedin 112 ml of absolute methylene chloride at 0° C. After addition of 56ml of anisole and 56 ml of trifluoroacetic acid, the mixture is stirredat 0° C. for 25 minutes. It is concentrated in vacuo, 100 ml of benzeneare added and the batch is stirred under a high vacuum for 1 hour. Theresidue is dissolved in 100 ml of absolute tetrahydrofuran, and 18.6 g(188 mmol) of N-ethylpyrrolidine are added. The solution is stirred atroom temperature for 30 minutes. 100 ml of ether are added. Theprecipitate formed is filtered off with suction, washed with 500 ml ofether and dissolved in 50 ml of water by adding NaHCO₃. 4 g ofimmobilized penicillin G acylase are then added and the pH value is keptconstant at 7.8 by addition of 4N triethylamine in ethanol. When theenzymatic splitting has ended, the acylase is filtered off and thefiltrate is brought to pH 2 with concentrated hydrochloric acid. Theprecipitate formed is filtered off with suction over silica gel and thefiltrate is added dropwise to 2 l of acetone. The desired productcrystallises out as the hydrochloride and is filtered off with suctionand dried.

Yield: 1.76 g (×HCl×H₂ O, 25.6%).

NMR (D₂ O): δ(ppm)=5.31 (1H, d, J=5 Hz, H-7-lactam); 5.12 (1H, d, J=5Hz, H-6-lactam); 4.62 (1H, d, J=14 Hz, CH₂ -pyrrolidine); 3.88 (1H, d,J=14 Hz, CH₂ -pyrrolidine); 3.86 (1H, d, J=18 Hz, S-CH₂); 3.58 (1H, d,J=18 Hz, S--CH₂); 3.42 (4H, m, pyrrolidine); 3.24 (2H, q, J=7 Hz, --CH₂--N⁺ --); 2.06 (4H, m, pyrrolidine); and 1.24 (3H, t, J=7 Hz, CH₃).

EXAMPLE 37β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-(1-ethyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate

353 mg (1.76 mmol) of (Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoaceticacid are dissolved in 2.7 ml of absolute dimethylformamide undernitrogen at room temperature. After addition of 113 μl ofN-ethyldiisopropylamine, 123 μl of tripropylamine and 152 μl oftributylamine, the mixture is cooled to -50° C. 145 μl ofmethanesulphonyl chloride are added and the solution is stirred at -50°C. for 30 minutes. This solution is then added rapidly to a solution,cooled to 0° C., of 470 mg (1.35 mmol) of7-amino-3-(1-ethyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate (×HCl)in 0.85 ml of water and 0.7 ml of triethylamine. After 5 minutes, thereaction solution is added to 150 ml of acetone. The precipitate formedis filtered off with suction, dried and chromatographed over adsorberresin HP 20 (eluting agent: acetonitrile/water 5/95).

Yield: 400 mg (59%)

¹ H-NMR (D₆ -DMSO): δ(ppm)=9.63 (1H, d, J=9 Hz, NH); 7.28 (2H, bs, NH₂);6.78 (1H, s, thiazole); 5.69 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.15(1H, d, J=5 Hz, H-6-lactam); 5.08 (1H, d, J=14 Hz, CH₂ -pyrrolidine);3.88 (1H, s, OCH₃); 3.85 (1H, d, J=14 Hz, CH₂ -pyrrolidine); 3.81 (1H,d, J=18 Hz, S--CH₂); 3.20-3.50 (7H, m, S--CH₂, --CH₂ --N--,pyrrolidine); 2.04 (4H, m, pyrrolidine); and 1.28 (3H, t, J=7 Hz, CH₃).

EXAMPLE 47-Amino-3-(1-ethyl-1-piperidinium)methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 2 from benzhydryl3-chloromethyl-7β-phenylacetamido-3-cephem-4-carboxylate andN-ethylpiperidine.

¹ H-NMR (D₂ O): δ(ppm)=5.31 (1H, d, J=5 Hz, H-7-lactam); 5.10 (1H, d,J=5 Hz, H-6-lactam); 4.59 (1H, d, J=15 Hz, CH₂ -piperidine); 3.89 (1H,d, J=18 Hz, S--CH₂); 3.87 (1H, d, J=15 Hz, CH₂ -piperidine); 3.45 (1H,d, J=18 Hz, S--CH₂); 3.34 (2H, q, J=7 Hz, --CH₂ +N--); 3.00-3.20 (4H, m,piperidine); 1.40-1.70 (6H, m, piperidine); and 1.19 (3H, t, J=7 Hz,CH₃).

EXAMPLE 57β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-(1-ethyl-1-piperidinium)methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-(1-ethyl-1-piperidinium)methyl-3-cephem-4-carboxylate.

¹ H-NMR (D₆ -DMSO): δ(ppm)=9.65 (1H, d, J=9 Hz, NH); 7.26 (2H, bs, NH₂);6.77 (1H, s, thiazole); 5.70 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.13(1H, d, J=5 Hz, H-6-lactam); 5.10 (1H, d, J=14 Hz, CH₂ -piperidine);3.80-3.90 (5H, m, OCH₃, CH₂ -piperidine, S--CH₂); 3.30-3.50 (7H, m,S--CH₂ --N--, piperidine); 1.40-1.80 (6H, m, piperidine); and 1.19 (3H,t, J=7 Hz, CH₃).

EXAMPLE 63-[(1-2-Hydroxyethyl)-1-pyrrolidinium]methyl-7β-phenylacetamido-3-cephem-4-carboxylate

4.68 g (12 mmol) of3-acetoxymethyl-7β-phenylacetamido-3-cephem-4-carboxylic acid aresuspended in 48 ml of absolute methylene chloride under nitrogen at roomtemperature and are dissolved by addition of 7.6 ml (36 mmol)N-methyl-N-trimethyl-silyltrifluoroacetamide (MSTFA). After cooling to0° C., 7 ml (48 mmol) of trimethylsilyl iodide are added and thereaction solution is stirred at 0° C. for 1 hour. After addition of 7.6ml of absolute tetrahydrofuran, the mixture is stirred at 0° C. for afurther 15 minutes. 14.4 ml (120 mmol) of N-(2-hydroxyethyl)pyrrolidineare then added and the solution is subsequently stirred for 30 minutes.2.4 ml of water are then added and, after a further 5 minutes, themixture is poured onto 200 ml of ether. The ether is decanted off fromthe oily residue, the residue is stirred again with ether and, afterdecanting again, is taken up in water, and the mixture ischromatographed over adsorber resin HP 20 (eluting agent:acetonitrile/water 5/95).

Yield: 3.6 g (68%).

¹ H-NMR (D₆ -DMSO): δ(ppm)=9.13 (1H, d, J=9 Hz, NH); 7.28 (5H, m,aromatic); 5.55 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.06 (1H, d, j=5Hz, H-6-lactam); 5.04 (1H, d, J=14 Hz, CH₂ -pyrrolidine); 3.95 (1H, d,J=14 Hz, CH₂ -pyrrolidine); 3.33-3.85 (12H, m); and 2.04 (4H, m,pyrrolidine).

EXAMPLE 77-Amino-3-[1-(2-hydroxyethyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate

4 g of immobilized penicillin G acylase are added to a solution of 3.5 g(7.8 mmol) of3-[1-(2-hydroxyethyl-1-pyrrolidinium]methyl-7β-phenylacetamido-3-cephem-4-carboxylatein 100 ml of water and the pH value is kept constant at 7.8 by adding 4Ntriethylamine in ethanol. When the enzymatic splitting has ended, theacylase is filtered off and the filtrate is brought to pH 2 withconcentrated hydrochloric acid. The precipitate formed is filtered offwith suction over silica gel and the filtrate is added dropwise to 2 lof acetone. the desired product crystallizes out as the hydrochlorideand is filtered off with suction and dried.

Yield: 1.9 g (×HCl×H₂ O, 64%).

¹ H-NMR (D₆ -DMSO): δ(ppm)=5.33 (1H, d, J=5 Hz, H-7-lactam); 5.13 (1H,d, J=5 Hz, H-6-lactam); 4.70 (1H, d, J=14 Hz, CH₂ -pyrrolidine); 3.93(2H, m, CH₂ --OH); 3.87 (1H, d, J=18 Hz, S--CH₂); 3.30-3.70 (7H, m); and2.11 (4H, m, pyrrolidine).

EXAMPLE 87β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-[1-(2-hydroxyethyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate.

¹ H-NMR (D₆ -DMSO): δ(ppm)=9.58 (1H, d, J=) Hz, NH); 9.24 (2H, bs, NH₂);6.71 (1H, s, thiazole); 5.64 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.11(1H, d, J=5 Hz, H-6-lactam); 5.03 (1H, d, J=13 Hz, CH₂ -pyrrolidine);3.93 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 3.81 (3H, s, OCH₃); 3.80 (2H,m, CH₂ --OH); 3.77 (1H, d, J=18 Hz, S--CH₂); 3.30-3.60 (7H, m); and 2.01(4H, m, pyrrolidine).

EXAMPLE 93-[1-(2-Hydroxyethyl)-1-piperidinium]methyl-7β-phenylacetamido-3-cephem-4-carboxylate

This compound is prepared analogously to Example 6 from3-acetoxymethyl-7β-phenylacetamido-3-cephem-4-carboxylic acid andN-(2-hydroxyethyl)-piperidine.

¹ H-NMR (D₆ -DMSO): δ(ppm)=9.17 (1H, d, J=9 Hz, NH); 7.30 (5H, m,aromatic); 5.56 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.09 (1H, d, J=5Hz, H-6-lactam); 5.08 (1H, d, J=13 Hz, CH₂ -piperidine); 3.10-3.90 (12H,m); and 1.40-1.90 (6H, m).

EXAMPLE 107-Amino-3-[1-(2-hydroxyethyl)-1-piperidinium]methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 7 from3-[1-(2-hydroxyethyl)-1-piperidinium]methyl-7β-phenylacetamido-3-cephem-4-carboxylate.

¹ H-NMR (D₂ O): δ(ppm)=5.34 (1H, d, J=5 Hz, H-7-lactam); 5.14 (1H, d,J=5 Hz, H-6-lactam); 4.75 (1H, d, J=14 Hz, CH₂ -piperidine); 3.96 (2H,m, CH₂ -OH); 3.91 (1H, d, J=18 Hz, S-CH₂); 3.10-3.60 (7H, m); and1.40-1.90 (6H, m, piperidine).

EXAMPLE 117β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoactamido]-3-[1-(2-hydroxyethyl)-1-piperidinium]methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-[1-(2-hydroxyethyl)-1-piperidinium]methyl-3-cephem-4-carboxylate.

¹ H-NMR (D₆ -DMSO): δ(ppm)=9.61 (1H, d, J=9 Hz, NH); 7.26 (2H, bs, NH₂);6.74 (1H, s, thiazole); 5.67 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.15(1H, d, J=5 Hz, H-6-lactam); 5.09 (1H, d, J=14 Hz, CH₂ -piperidine);4.01 (1H, d, J=14 Hz, CH₂ piperidine); 3.84 (3H, s, OCH₃); 3.80 (3H, m);3.10-3.50 (7H, m); and 1.40-1.90 (6H, m, piperidine).

EXAMPLE 123-[4-(2-Hydroxyethyl)-4-morpholinium]methyl-7β-phenylacetamido-3-cephem-4-carboxylate

This compound is prepared analogously to Example 6 from3-acetoxymethyl-7β-phenylacetamido-3-cephem-4-carboxylic acid andN-(2-hydroxyethyl)morpholine.

¹ H-NMR (DMSO-D₆): δ(ppm)=9.19 (1H, d, J=9 Hz, NH); 7.34 (5H, m,aromatic); 5.62 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.20 (1H, d, J=14Hz, CH₂ -morpholine); 5.13 (1H, d, J=5 Hz, H-6-lactam); 4.16 (1H, d,J=14 Hz, CH₂ -morpholine); and 3.30-4.10 (16H, m).

EXAMPLE 137-Amino-3-[4-(2-hydroxyethyl)-4-morpholinium]methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 7 from3-[4-(2-hydroxyethyl)-4-morpholinium]methyl-7β-phenylacetamido-3-cephem-4-carboxylate.

¹ H-NMR (D₂ O): δ(ppm)=5.36 (1H, d, J=5 Hz, H-7-lactam); 5.15 (1H, d,J=5 Hz, H-6-lactam); 4.88 (1H, d, J=14 Hz, CH₂ -morpholine); 4.21 (1H,d, J=14 Hz, CH₂ -morpholine); and 3.30-4.10 (14H, m).

EXAMPLE 147β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[4-(2-hydroxyethyl)-4-morpholinium]methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-[4-(2-hydroxyethyl)-4-morpholinium]methyl-3-cephem-carboxylate.

¹ NMR (DMSO-D₆): δ(ppm)=9.63 (1H, d, J=9 Hz, NH); 7.28 (2H, bs, NH₂);6.77 (1H, s, thiazole); 5.71 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.21(1H, d, J=14 Hz, CH₂ -morpholine); 5.11 (1H, d, J=5 Hz, H-6-lactam);4.13 (1H, d, J=14 Hz, CH₂ -morpholine); 3.87 (3H, s, OCH₃); and3.30-4.00 (14H, m).

EXAMPLE 157-Amino-3-(4-ethyl-4-morpholinium)methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 17 from benzhydryl3-chloromethyl-7β-phenylacetamdio-3-cephem-4-carboxylate.

¹ H-NMR (DMSO-d₆): δ(ppm)=5.35 (1H, d, J=5 Hz, H-7-lactam); 5.16 (1H, d,J=5 Hz, H-6-lactam); 4.58 (1H, d, J=14 Hz, CH₂ -morpholine); 4.02 (1H,d, J=14 Hz, CH₂ morpholine); 3.96 (4H, m, morpholine); 3.90 (1H, d, J=18Hz, S-CH₂); 3.30-3.60 (7H, m, S-CH₂, morpholine, ##STR21## and 1.24 (3H,t, J=7 Hz, CH₃).

EXAMPLE 167β[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-(4-ethyl-4-morpholinium)methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-(4-ethyl-4-morpholinium)methyl-3-cephem-4-carboxylate

¹ H-NMR (DMSO-d₆): δ(ppm)=9.59 (1H, d, J=9 Hz, NH); 7.25 (2H, bs, NH₂);6.76 (1H, s, thiazole); 5.69 (1H, dd, J=9Hz, J=5 Hz, H-7-lactam); 5.23(1H, d, J=13 Hz, CH₂ -morpholine); 5.16 (1H, d, J=5 Hz, H-6-lactam),3.80-4.00 (6H, m, CH₂ -morpholine, morpholine, S-CH₂); 3.86 (3H, s,OCH₃); 3.30-3.60 (7H, m, S-CH₂, morpholine, --CH₂ --N--); and 1.25 (3H,t, J=7 Hz, CH₃).

EXAMPLE 177-Amino-3-(1-propyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 17 from benzhydryl3-chloromethyl-7β-phenylacetamido-3-cephem-4-carboxylate andN-propylpyrrolidine.

¹ H-NMR (D₂ O): δ(ppm)=5.33 (1H, d, J=5 Hz, H-7-lactam); 5.15 (1h, d,J=5 Hz, H-6-lactam); 4.63 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 3.96 (1H,d, J=13 Hz, CH₂ -pyrrolidine); 3.88 (1H, d, J=18 Hz, S-CH₂); 3.55 (1H,d, J=18 Hz, S-CH₂); 3.45 (4H, m, pyrrolidine); 3.12 (2H, m, ##STR22##2.10 (4H, m, pyrrolidine; 1.66 (2H, m, --CH₂); and 0.85 (3H, m, CH₃).

EXAMPLE 187β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-(1-propyl-1-pyrrolidinium)methyl)-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-(1-propyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate.

¹ H-NMR (DMSO-d₆): δ(ppm)=9.58 (1H, d, J=9 Hz, NH); 7.24 (2H, bs, NH₂);6.75 (1H, s, thiazole); 5.67 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.13(1H, d, J=5 Hz, H-6-lactam); 5.08 (1H, d, J=13 Hz, CH₂ -pyrrolidine);3.86 (3H, s, OCH₃); 3.80 (2H, m, CH₂ -pyrrolidine, S--CH₂); 3.45 (5H, m,pyrrolidine, S--CH₂); 3.13 (2H, m, ##STR23## 2.03 (4H, m, pyrrolidine);1.74 (2H, m, --CH₂ --); and 0.90 (3H, t, J=7 Hz, CH₃).

EXAMPLE 197-Amino-3-(1-isopropyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 2 from benzhydryl3-chloromethyl-7β-phenylacetamido-3-cephem-4-carboxylate andN-isopropylpyrrolidine.

¹ H-NMR (D₂ O): δ (ppm)=5.35 (1H, d, J=5 Hz, H-7-lactam); 5.13 (1H, d,J=5 Hz, H-6-lactam); 4.02 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 3.93 (1H,d, J=18 Hz, S--CH₂); 3.40-3.80 (6H, m, S--CH₂, pyrrolidine, ##STR24##2.10 (4H, m, pyrrolidine); and 1.43 (6H, m, isopropyl).

EXAMPLE 207β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-(1-isopropyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-(1-isopropyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate.

¹ H-NMR (DMSO-d₆): δ (ppm)=9.61 (1H, d, J=9 Hz, NH); 7.28 (2H, s, NH₂);6.77 (1H, s, thiazole); 5.67 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.17(1H, d, J=5 Hz, H-6-lactam); 4.96 (1H, d, J=13 Hz, CH₂ -pyrrolidine);3.92 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 3.87 (3H, s, OCH₃); 3.83 (1H,d, J=18 Hz, S--CH₂); 3.40-3.70 (6H, m, ##STR25## pyrrolidine, S--CH₂);1.98 (4H, m, pyrrolidine); and 1.35 (6H, m, isopropyl).

EXAMPLE 217-Amino-3-(1-butyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 2 from benzhydryl3-chloromethyl-7β-phenylacetamido-3-cephem-4-carboxylate andN-butyl-pyrrolidine.

¹ H-NMR(D₂ O): δ (ppm)=5.24 (1H, d, J=5 Hz, H-7-lactam); 5.04 (1H, d,J=5 Hz, H-6-lactam); 4.58 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 3.85 (1H,d, J=13 Hz, CH₂ -pyrrolidine); 3.82 (1H, d, J=18 Hz, S--CH₂); 3.43 (1H,d, J=18 Hz, S--CH₂); 3.28 (4H, m, pyrrolidine); 3.07 (2H, m, ##STR26##2.04 (4H, m, pyrrolidine); 1.58 (2H, m, --CH₂ --); 1.18 (2H, m, --CH₂--); and 0.80 (3H, t, J=7 Hz, CH₃).

EXAMPLE 227β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-(1-butyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-(1-butyl-1-pyrrolidinium)methyl-3-cephem-4-carboxylate.

¹ H-NMR (DMSO-d₆): δ (ppm)=9.58 (1H, d, J=9 Hz, NH); 7.21 (1H, bs, NH₂);6.71 (1H, s, thiazole); 5.62 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.10(1H, d, J=5 Hz, H-6-lactam); 5.05 1H, d, J=13 Hz, CH₂ -pyrrolidine);3.86 (3H, s, OCH₃); 3.82 (2H, m, CH₂ -pyrrolidine, S--CH₂); 3.30 (5H, m,S--CH₂, pyrrolidine); 3.13 (2H, m, ##STR27## 2.01 (4H, m, pyrrolidine);1.70 (2H, m, --CH₂ --); 1.27 (2H, m, --CH₂); and 0.90 (3H, t, J=7 Hz,CH₃).

EXAMPLE 233-[1-(3-Hydroxypropyl)-1-pyrrolidinium]methyl-7β-phenylacetamido-3-cephem-4-carboxylate

This compound is prepared analogously to Example 6 from3-acetoxymethyl-7β-phenylacetamido-3-cephem-4-carboxylic acid andN-(3-hydroxypropyl)-pyrrolidine.

¹ H-NMR (D₆ -DMSO): δ (ppm)=9.16 (1H, d, J=9 Hz, NH); 7.30 (5H,aromatic); 5.58 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.08 (1H, d, J=5Hz, H-6-lactam); 5.03 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 3.88 (1H, d,J=13 Hz, CH₂ -pyrrolidine); 3.84 (1H, d, J=18 Hz, S--CH₂); 2.90-3.60(11H, m); and 1.80-2.10 (6H, m).

EXAMPLE 247-Amino-3-[1-(3-hydroxypropyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 7 from3-[1-(3-hydroxypropyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate.

¹ H-NMR(D₂ O): δ (ppm)=5.31 (1H, d, J=5 Hz, H-7-lactam); 5.11 (1H, d,J=5 Hz, H-6-lactam); 4.63 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 3.96 (1H,d, J=13 Hz, CH₂ -pyrrolidine); 3.88 (1H, d, J=18 Hz, S--CH₂); 3.40-3.60(9H, m); 2.08 (4H, m, pyrrolidine); and 1.92 (2H, m, --CH₂ --).

EXAMPLE 257β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-(3-hydroxypropyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-[1-(3-hydroxypropyl)1-pyrrolidinium]methyl-3-cephem-4-carboxylate.

¹ H-NMR (DMSO-d₆): (ppm)=9.59 (1H, d, J=9 Hz, NH); 7.26 (2H, bs, NH₂);6.75 (1H, s, thiazole); 5.68 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.13(1H, d, J=5 Hz, H-6-lactam); 5.05 (1H, d, J=13 Hz, CH₂ -pyrrolidine);3.86 (3H, s, OCH₃); 3.84 (2H, m, CH₂ -pyrrolidine, S--CH₂); 3.20-3.60(9H, m, S--CH₂, --CH₂ --N--, --CH₂ OH' pyrrolidine); and 1.88 (2H, m,--CH₂ --).

EXAMPLE 263-{1-[2-(2-Hydroxyethoxy)ethyl]-1-pyrrolidinium}methyl-7β-phenylacetamido-3-cephem-4-carboxylate

This compound is prepared analogously to Example 6 from3-acetoxymethyl-7β-phenylacetamido-3-cephem-4-carboxylic acid andN-[2-(2-hydroxyethoxy)ethyl]-pyrrolidine.

¹ H-NMR (D₆ -DMSO): δ (ppm)=9.15 (1H, d, J=9 Hz, NH); 7.31 (5H, m,aromatic); 5.57 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.15 (1H, d, J=13Hz, CH₂ -pyrrolidine); 5.08 (1H, d, J=5 Hz, H-6-lactam); 3.99 (1H, d,J=13 Hz, CH₂ -pyrrolidine); 2.90-3.90 (14H, m); and 2.08 (4H, m,pyrrolidine).

EXAMPLE 277-Amino-3-{1-[2-(2-hydroxyethoxy)ethyl]-1-pyrrolidinium}methyl-3-cephem-4-carboxylate

This compund is prepared analogously to Example 7 from3-{1-[2-(2-hydroxyethoxy)ethyl]-1-pyrrolidinium}methyl-7β-phenylacetamido-3-cephem-4-carboxylate.¹ H-NMR (D₂ O): δ (ppm)=5.25 (1H, d, J=5 Hz, H-7-lactam); 5.07 (1H, d,J=5 Hz, H-6-lactam); 4.65 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 4.04 (1H,d, J=13 Hz, CH₂ -pyrrolidine); 3.82 (1H, d, J=18 Hz, S--CH₂); 3.30-380(9H, m); and 2.05 (4H, m, pyrrolidine).

EXAMPLE 287β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-{1-[2-(2-hydroxyethoxy)ethyl]1-pyrrolidInium}methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-{1-[2-(2-hydroxyethoxy)ethyl]-1-pyrrolidinium}methyl-3-cephem-4-carboxylate.

¹ H-NMR (DMSO-d₆): δ (ppm)=9.60 (1H, d, J=9 Hz, NH); 7.27 (2H, bs, NH₂);6.74 (1H, s, thiazole); 5.67 (1H, dd, J=9 Hz, J=5 Hz, H-7-lactam); 5.16(1H, d, J=13 Hz, CH₂ -pyrrolidine); 5.14 (1H, d, J=5 Hz, H-6-lactam);3.98 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 3.84 (3H, s, OCH₃); 3.80 (3H,m); 3.30-3.60 (11H, m); and 2.05 (4H, m, pyrrolidine).

EXAMPLE 293-[1-(2-Hydroxy-2-phenylethyl)1-pyrrolidinium]methyl-7β-phenylacetamido-3-cephem-4-carboxylate

This compound is prepared analogously to Example 6 from3-acetoxymethyl-7β-phenylacetamido-3-cephem-4-carboxylic acid andDL-N-(2-hydroxy-2-phenylethyl)pyrrolidine. A mixture of twodiastereoisomers is obtained.

¹ H-NMR (D₆ -DMSO): δ (ppm)=9.14 (1H, d, J=9 Hz, NH); 7.20-7.60 (10H, m,aromatic); 5.58 (1H, m H-7-lactam); 5.33 (1H, m, CH--OH); 5.19 (1H, m,CH₂ -pyrrolidine); 5.07 (1H, d, J=5 Hz, H-6-lactam); 4.36 and 4.21 (1H,d, J=13 Hz, CH₂ -pyrrolidine); 3.10-390 (10H, m); and 2.10 (4H, m,pyrrolidine).

EXAMPLE 307-Amino-3-[1-(2-hydroxy-2-phenylethyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 7 from3-[1-(2-hydroxy-2-phenylethyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate(mixture of two diastereoisomers).

¹ H-NMR (D₂ O): δ (ppm)=7.40 (5H, bs, aromatic); 5.30 (2H, m,H-7-lactam, CH--OH); 5.14 (1H, m H-6-lactam); 4.82 (1H, m, CH₂-pyrrolidine); 4.55 and 4.37 (1H, d, J=13 Hz, CH₂ -pyrrolidine);3.30-4.00 (8H, m); and 2.16 (4H, bs, pyrrolidine).

EXAMPLE 317β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-(2-hydroxy-2-phenylethyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate

This compound is prepared analogously to Example 3 from(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetic acid and7-amino-3-[1-(2-hydroxy-2-phenylethyl)-1-pyrrolidinium]methyl-3-cephem-4-carboxylate(mixture of two diastereoisomers).

¹ H-NMR (DMSO-d₆): δ (ppm)=9.58 (1H, d, J=9 Hz, NH); 7.30-7.50 5H, m,aromatic); 7.22 (2H, bs, NH₂); 6.72 and 6.73 (1H, s, thiazole); 5.63(1H, m, H-7-lactam); 5.08-5.30 (3H, CH--OH, CH₂ -pyrrolidine,H-6-lactam); 4.29 and 4.13 (1H, d, J=13 Hz, CH₂ -pyrrolidine); 3.82 (3H,s, OCH₃); 3.20-3.80 (8H, m; and 2.05 (4H, m, pyrrolidine).

It will be understood that the specification and examples areillustrative but not limitative of the present invention and that otherembodiments within the spirit and scope of the invention will suggestthemselves to those skilled in the art.

We claim:
 1. A cephalosporin which is selected from the group consistingof7β-[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-(1-ethyl-1-piperidinium)methyl-3-cephem-4-carboxylateand7β[(Z)-2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-(4-ethyl-4-morpholinium)methyl-3-cephem-4-carboxylateor a pharmaceutically acceptable salt thereof.
 2. An antibacterialcomposition comprising an antibacterially effective amount of a compoundor salt according to claim 1 and a diluent.
 3. A unit dose of acomposition according to claim 1, in the form of a tablet, capsule orampule.
 4. A method of combatting bacteria which comprises applying tosuch bacteria or to a bacteria habitat an antibacterially effectiveamount of a cephalosporin or salt according to claim
 1. 5. An animalgrowth promoting composition comprising an animal growthpromoting-effective amount of a compound or salt according to claim 1and an animal foodstuff as a diluent.